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After ozone therapy for bleaching, it is important to evaluate enamel surface properties, to ensure that bleaching provides adequate conditions for sound dental substrate. Aim: The aim of this in vitro study was to evaluate the effects of a bleaching treatment with 10% carbamide peroxide (CP), with or without ozone (O), on the microhardness, roughness and micromorphology of the enamel surface., Materials and Method: Bovine enamel blocks were planed and distributed among the following three bleaching treatment groups (n=10): CP - 1 hour per day/14 days (Opalescence PF 10%/ Ultradent); O - 1 hour per day every 3 days/3 sessions (Medplus V Philozon, 60 mcg/mL and oxygen flow rate of 1 L/min); and OCP - CP with O, 1 hour per day every 3 days/3 sessions. Enamel surface microhardness (Knoop), roughness (Ra), and micromorphology by scanning electron microscopy (5,000x magnification) were determined before and after the treatments., Results: ANOVA and Tukey-Kramer’s test showed that enamel microhardness remained unchanged by treatment with O and OCP (p=0.0087), but decreased by treatment with CP. Treatment with O promoted higher enamel microhardness than the other groups (p=0.0169). Generalized linear mixed models for repeated measures over time indicated treatment with CP increased enamel roughness more than OCP or O (p=0.0003). CP produced slight irregularities in enamel micromorphology after the whitening treatment. O, with or without CP, maintained the mechanical and physical properties of microhardness and enamel surface micromorphology, and either maintained or reduced surface roughness, compared to the conventional tray-delivered CP bleaching treatment., Conclusions: Treatment with 10% carbamide peroxide in trays promoted greater changes in enamel surface properties than treatments with ozone and with 10% ozonized carbamide peroxide in the office., Competing Interests: The authors declare no potential conflicts of interest regarding the research, authorship, and/or publication of this article.